Sexual dimorphism in ultradian and 24h rhythms in plasma levels of growth hormone in Indian walking catfish, Clarias batrachus.

Growth hormone (GH), a key regulator of somatic and reproductive growth in vertebrates, has been extensively studied, although primarily in female fish. Despite numerous reports about sex- and species-specific growth patterns in fish, to our knowledge, there is no report about the 24 h rhythm of plasma GH in male fish. Thus, we aimed to investigate temporal variations in plasma GH levels and the existence of any rhythms therein during the reproductively active months of March to August in the male walking catfish, Clarias batrachus. We also aimed to compare the secretory temporal patterns of GH in male-female specimens of C. batrachus to decipher sexual dimorphism in GH secretions in fish. After 14 days of acclimation to the natural environment, male catfish (N = 240 in total) were sorted and randomly divided into eight groups for study at ZT0 (sunrise ~06:00 h), 3, 6, 9, 12, 15, 18, and 21. During each month, physical parameters like duration of photoperiod and water temperature were measured. Male catfish (n = 40/month) in all eight groups were sampled (n = 5/group) at each time point under the natural time-of-year 24 h light-dark (LD) cycle. Male catfish were anesthetized and blood was collected through a caudal puncture, centrifuged, and plasma isolated. Plasma GH was measured using a competitive homologous enzyme-linked immunosorbent assay. Further, testes were removed, weighed, and the gonadosomatic index (GSI) was calculated. A significant effect of time and season (p ˂ 0.05, two-way ANOVA) on plasma GH level was detected. Cosinor analyses verified the existence of statistically significant (p ˂ 0.05) ultradian (12 h) and 24 h rhythms of plasma GH in male C. batrachus, with the higher values of Mesor (time series mean) and amplitude (one-half peak-to-trough difference) of the periodicities from March to July. Mapping of the acrophases (peak times) showed two ultradian and one 24 h acrophase of GH during the early photophase and early scotophase from March to August. Distinct sexual-dimorphism in plasma GH Mesors and acrophases was noticed between male and female catfish. GSI values of male catfish indicate males mature a little earlier than females in terms of size and reproductive activity. The findings that plasma GH show 24 h and seasonal fluctuations in a sex-specific manner collectively demonstrate the importance of considering the effect of biological 24 h and seasonal time and sex on the GH level in regulating the physiology of somatic growth and reproduction in catfish.

Localization, distribution and expression of growth hormone in the brain of Asian Catfish, Clarias batrachus.

Intense immunoreactivity was observed in several neurons of the nucleus preopticus (NPO) located in the preoptic area (POA), in addition to several GH cells in the proximal pars distalis (PPD) of the pituitary gland of Clarias batrachus. The immunoreactive cells were located in the paraventricular as well as supraoptic subdivisions of the NPO. GH immunoreactive fibers projecting from the neurons were traced caudally to the pituitary gland via the conspicuous preoptico-hypophysial tract (PHT) in the ventral tuberal area to the neurohypophysis of the pituitary. Apart from these immunoreactive fibers in the preoptico-hypophysial tract, some fine caliber fiber probably arising from the neurons located dorsally in the NPO also showed GH immunoreactivity, and these fibers constituted an independent tract. Bilaterally, it extended caudally through the dorsal hypothalamus almost as far as the saccus vasculosus where it curved sharply to descend into the caudal tuberal region and finally entered into the pituitary gland. The fibers of this tract were mainly distributed in the rostral pars distalis (RPD). This tract is quite distinct from the preoptico-hypophysial tract and herein called as the accessory preoptico-hypophysial tract (APHT). Expression of GH mRNA in the NPO was found 65-fold more than that of the control region, rostral cerebellum. These results altogether suggest that GH secreted by NPO neurons might serve as a neuro-modulatory role in the brain of C. batrachus. Transportation of GH to the pituitary via two independent tracts may represent the duality of neuroendocrine function. The present study underscores the possibility that GH in the brain of vertebrates may be a phylogenetically conserved phenomenon and provide clues to our understanding of the evolutionary course taken by the hormone.

Molecular analysis of dinucleotide microsatellite in growth hormone gene of Asian seabass (Lates calcarifer) from Mumbal, India.

In the present study, out of four alleles amplified from seabass (Lates calcarifer) genome inhabiting Mumbai water by PCR using growth hormone (GH) gene-specific primers, two DNA fragments (SGMS1, 233 bp and SGMS2, 239 bp) were eluted from gel, cloned using pTZ57R (2.886 kb) vector into E. coli DH5α, characterized by restriction endonuclease analysis and sequenced by automated DNA sequencer. After blasting and multiple alignment of the above sequences, SGMS1 showed 97% and SGMS2 93.3% homology with promoter region of GH gene containing microsatellite of Australian seabass and 94.6% homology between both the fragments. These sequences SGMS1 and SGMS2 were submitted to NCBI GenBank. On blasting, these sequences with gene databases, SGMS1 and SGMS2 showed partial homologies with Seriola quinqueradiata (26.9%, 12.9%), flounder (15.8%, 15.8%), Oreochromis nilotica (23%, 7.9%), Oreochromis mossambicus (23%, 7.9%) and Danio rerio (8.2%, 7.5%). Critical analysis showed the presence of microsatellite (CA)16 and (CA)19 repeats in fragments SGMS1 and SGMS2, respectively in seabass from Mumbai water in comparison to (CA)14 repeats from the Australian seabass. Further, on sequence comparison, single nucleotide mismatches detected at their several positions in relation to seabass GH gene of Australia. These nucleotide variations detected in SGMS1 and SGMS2 in comparison to those of the Australian seabass may be due to mutations owing to environmental or habitat changes that seem to have definite potentials for development of genetic markers, which would be useful for identification and selection of superior germplasm with desirable commercial traits such as high growth rate.

Temperature dependent action of growth hormone on somatic growth and testicular activities of the catfish, Clarias batrachus.

Effects of growth hormone on somatic growth and testicular activities were studied during late quiescence and early recrudescence phases of the reproductive cycle of the catfish, Clarias batrachus. The administration of exogenous growth hormone (GH) during the late quiescence phase (December-January; ambient water temperature-15.2±1°C) did not influence the somatic growth as well as the testicular activity, as no change in body weight, testis weight, plasma level of insulin-like growth factor I (IGF-I) and testicular morphology was detected following GH treatment, though the plasma testosterone was marginally increased. While during the early recrudescence phase (March-April; ambient water temperature-28.1±2°C), GH treatment promoted the production of insulin like growth factor-I and testicular steroidogenic activity in a dose dependent manner, as was evident from the significant increase in the circulating levels of testosterone and estradiol-17ß. GH treatment also increased body weight, testicular weight and gonadosomatic index, suggesting its involvement in testicular development. The GH treatment promoted spermatogonial proliferation and accelerated the spermatogenic process in the present catfish. These results, thus, suggest that GH influences the somatic growth and testicular activities depending on the temperature of the rearing water; warmer temperature and longer photoperiod promote testicular steroidogenic and spermatogenic activities in fish. This study has immense practical use in fisheries science.